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  4. 新药研究国家重点实验室
High-affinity nanobodies targeting IL-12B for the detection of fluorescence resonance energy transfer

文献类型：期刊论文

作者Hu, Jing3; Feng, Wenxuan3; Wen, Jianchuan1; Zhou, Siyu2; Sun, Zengchao4,5; Zhao, Shuaiying1; Guo, Shaojue1; Wang, Hui1; Geng, Yong2,3
刊名PROTEIN EXPRESSION AND PURIFICATION
出版日期2025-05-01
卷号229页码:9
关键词Nanobody screening IL-12B Phage display technology Fluorescence resonance energy transfer
ISSN号1046-5928
DOI10.1016/j.pep.2025.106681
英文摘要Aims: IL-12B, a subunit of the IL-23 family of cytokines, plays a crucial role in various diseases such as viral infections, autoimmune disorders, and tumors. This study aimed to identify high-affinity nanobodies that bind to distinct epitopes of IL-12B to and assess their potential for therapeutic and diagnostic applications, particularly through fluorescence resonance energy transfer(FRET) to evaluate their ability to target IL-12B. Methods: IL-12B protein was expressed in eukaryotic cells and used to immunize camels to induce an immune response. Camel-derived anti-IL-12B nanobodies were isolated and screened via phage display to identify those with high specificity and affinity for IL-12B. Binding affinity and epitope interactions were further analyzed using high-performance liquid chromatography (HPLC) and ForteBio Octet assays. A FRET-based assay was developed to evaluate protein interactions for precise therapeutic targeting. Results: Several high-affinity nanobodies targeting IL-12B were successfully generated. These nanobodies exhibited strong binding to various epitopes of IL-12B. Screening by HPLC and ForteBio Octet confirmed their high specificity and affinity, while fluorescence analysis demonstrated efficient energy transfer between thenanobodies, indicating successful interactions. Conclusions: This study identified high-affinity nanobodies against IL-12B and used FRET to characterize their interactions. These nanobodies show promise for therapeutic potential targeting IL-12B-related diseases, including viral infections, autoimmune disorders, and cancer. However, further clinical studies are needed to fully explore their potential for diagnostic and therapeutic applications.
WOS关键词ANTIBODIES ; PROTEIN
WOS研究方向Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
语种英语
WOS记录号WOS:001440031400001
出版者ACADEMIC PRESS INC ELSEVIER SCIENCE
源URL[http://119.78.100.183/handle/2S10ELR8/316511]  
专题新药研究国家重点实验室
通讯作者Hu, Jing
作者单位1.Shanghai Ocean Univ, Coll Food Sci & Technol, Dept Biopharmaceut, Shanghai 201306, Peoples R China
2.Chinese Acad Sci, Shanghai Inst Mat Med, State Key Lab Drug Res, CAS Key Lab Receptor Res, Shanghai 201203, Peoples R China
3.Nanjing Univ Chinese Med, Sch Chinese Mat Med, Nanjing 210023, Jiangsu, Peoples R China
4.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai, Peoples R China
5.ShanghaiTech Univ, Shanghai Inst Adv Immunochem Studies, Shanghai, Peoples R China
推荐引用方式
GB/T 7714		 Hu, Jing,Feng, Wenxuan,Wen, Jianchuan,et al. High-affinity nanobodies targeting IL-12B for the detection of fluorescence resonance energy transfer[J]. PROTEIN EXPRESSION AND PURIFICATION,2025,229:9.
APA Hu, Jing.,Feng, Wenxuan.,Wen, Jianchuan.,Zhou, Siyu.,Sun, Zengchao.,...&Geng, Yong.(2025).High-affinity nanobodies targeting IL-12B for the detection of fluorescence resonance energy transfer.PROTEIN EXPRESSION AND PURIFICATION,229,9.
MLA Hu, Jing,et al."High-affinity nanobodies targeting IL-12B for the detection of fluorescence resonance energy transfer".PROTEIN EXPRESSION AND PURIFICATION 229(2025):9.

入库方式： OAI收割

来源：上海药物研究所

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